Patent GB-474822-A (GB)
Title: Process for the production of specifically active enzyme preparations
IPC: A21D6/00
Filed: 1936-02-01
Abstract: Enzyme preparations are obtained by treating micro-organisms containing several enzyme complexes, for example by drying, heating in media rich in albumen, so as to kill the cells while not destroying all the enzymic activity, and liberating "endo-enzymes," subjecting to low temperature, or treating with chemicals such as acetone or glycerin to kill the cells and remove certain enzymes such as lipase, while maintaining a residual activity of other enzymes such as proteinases and zymases.  The products may be mized with one another or with other enzyme materials (such as diastase, proteinase, or grain germs) and with substrates (such as carbohydrates and albumens, particularly milk and dried milk preparations) and activators such as organic acids (lactic acid and vinegar) materials containing co-zymases and organic substances containing phosphorus such as nucleo-proteids. The preparations may be used as baker's assistants for the production of bread, the production of malt, cheese, the rotting of manure, the retting of fibres, the fermentation of tobacco, cacao and coffee and other processes for the swelling and loosening of organic materials.  For the production of malt, the preparation should contain diastatic, proteolytic and zymatic properties, the cellulose and pectinase complexes being destroyed by means of bacteria or mould fungi which destroy cellulose and contain pectin.  Such preparations are suitable for rotting manure and retting fibres.  For the fermentation of tobacco the proteolytic enzyme is "selectively fortified."  For cheese-making the proteolytic enzyme is "selectively fortified" and enzymes containing lactase which may be obtained from milk-sugar yeast, may be added, such yeasts being living or partly destroyed while maintaining the lactase activity.  For the fermentation of cacao and coffee preparations are used in which the pectinases and germination ferment complexes are "selectively fortified" relatively to the other ferment complexes present.  As starting materials may be used bacteria rich in pectinase of the coli-aerogenes group and moulds rich in pectinase such as aspergillus; such preparations may be used together with material containing zymase and proteinase for instance from schizo-saccharomyces.  To the preparations may be added various specified materials for promoting or inhibiting the action of particular enzymes: tryptic enzymes may be inhibited by organic acids of pH at most 5; and for promoting zymase action magnesium sulphate sufficient to give a magnesium concentration of 0,01 to 0,03 per cent in the medium in which the preparation is to be used.  In examples: (1) a baker's assistant consists of 30 parts schizo-saccharomyces dried at 60--65 DEG  C. for at least a minute, preferably under reduced pressure, 10 to 15 parts of an aspergillus rich in diastase dried in a similar manner, 30 parts bran flour, 5 parts wheat germ, and 25 to 20 parts of dried milk. The schizo-saccharomyces bomb may be cultivated in known manner in a dilute beer wort free from hops, 5 per cent raw sugar with additional nutrients, or grape must culture medium.  The cultivation may be at 30 DEG  C. with aeration, and the organism may be separated from the medium, washed and dried as such or after dilution.  For the drying the pulp is supplied in a thin layer to a heated rotary drum with a stripper blade at one side, the vapour p  being withdrawn by a vacuum pump.  Instead of drying the pure yeast, the culture medium containing it may be dried.  The drying kills the cells, and inactivates the zymase and enzymes which break down low-molecular albuminous materials while proteinase is less affected.  In the cultivation of aspergillus it is important that no spores should be formed, as they are resistant to heat; (2) a baker's assistant consists of 40 parts of an autolyzed aspergillus of the torula, mycotorula, anomalus, mycoderma oidium and monilia groups, preferably those which are not rich in lipase; the organisms are cultivated in known manner, separated from the medium, killed by a suitable antiseptic such as chloroform or toluene and autolyzed at 40 DEG  C. and dried rapidly under reduced pressure; 15 parts of dried aspergillus, 10 parts wheat germ, 35 parts "excess from the preparation of the grain flour"; (3) for the fermentation of cacao the preparation contains 60 parts of aspergillus dried at 50--60 DEG C. while killing all the cells, and 40 parts of a preparation obtained by drying in the manner described above pectinase-containing bacteria of the coli-aerogenes group in which the cells are destroyed.  Preferably the preparation is used together with a pure yeast such as dako-schizo-saccharomyces.ALSO:Enzyme preparations are obtained by treating micro-organisms containing several enzyme complexes, for example by drying, heating in media rich in albumen so as to kill the cells while not destroying all the enzyme activity, and liberating "endo-enzymes," subjecting to low temperature, or treating with chemicals such as acetone or glycerin to kill the cells and remove certain enzymes such as lipase, while maintaining a residual activity of other enzymes such as proteinases and zymases. The products may be mixed with one another or with other enzyme materials (such as diastase, proteinase, or grain germs) and with substrates (such, as carbohydrates and albumens, particularly milk, and dried milk preparations) and, activators such as organic acids (lactic acid and vinegar) materials containing co-zymases and organic substances containing phosphorus such as nucleo-proteids.  The preparations may be used for the production of cheese, and the rotting of manure and other processes for the swelling and loosening of organic materials.  For the production of malt, the preparation should contain diastatic, proteolytic and zymatic properties, the cellulose and pectinase complexes being destroyed by means of bacteria or mould fungi which destroy cellulase and contain pectin. Such preparations are suitable for rotting manure and retting fibres. For cheese-making the proteolytic enzyme is "selectively fortified" and enzymes containing lactase which may be obtained from milk-sugar yeast, may be added, such yeast being living or partly destroyed while maintaining the lactase activity.  To the preparations may be added various specified materials for promoting or inhibiting the action of particular enzymes; tryptic enzymes may be inhibited by organic acids of pH at most 5; and for promoting zymase-action magnesium sulphate sufficient to give a magnesium concentration of 0,01 to 0,03 per cent in the medium in which the preparation is to be used.ALSO:Enzyme preparations are obtained by treating micro-organisms containing several enzyme complexes, for example by drying, heating in media rich in albumen, so as to kill the cells while not destroying all the enzyme activity, and liberating "endo-enzymes," subjecting to low temperature, or treating with chemicals such as acetone or glycerin to kill the cells and remove certain enzymes such as lipase, while maintaining a residual activity of other enzymes such as proteinases and zymases.  The products may be mixed with one another or with other enzyme materials (such as diastase, proteinase, or grain germs) and with substrates (such as carbohydrate and albumens, particularly milk, and dried milk preparations) and activators such as organic acids (lactic acid and vinegar) materials containing cozymases and organic substances containing phosphorus such as nucleo-proteids. The preparations may be used as bakers' assistants for the production of bread, the production of malt, the fermentation of tobacco, cacao and coffee and other processes for the swelling and loosening of organic materials. For the production of malt, the preparation should contain diastatic, proteolytic and zymatic properties, the cellulose and pectinase, complexes being destroyed by means of bacteria or mould fungi which destroy cellulase and contain pectin.  For the fermentation of tobacco the proteolytic enzyme is "selectively fortified."  For the fermentation of cacao and coffee preparations are used in which the pectinases and germination ferment complexes are "selectively fortified" relatively to the other ferment complexes present.  As starting materials may be used bacteria rich in pectinase of the coli-aerogenes group and moulds rich in pectinase such as aspergillus; such preparations may be used together with material containing zymase and proteinase, for instance from schizo-saccharomyces.  To the preparations may be added various specified materials for promoting or inhibiting the action of particular enzymes; tryptic enzymes may be inhibited by organic acids of pH at most 5; and for promoting zymase-action magnesium sulphate sufficient to give a magnesium concentration of 0,01 to 0,03 per cent in the medium in which the preparation is to be used.  In examples: (1) a baker's assistant consists of 30 parts schizo-saccharomyces dried at 60-65 DEG  C. for at least a minute, preferably under reduced pressure, 10 to 15 parts of an aspergillus rich in diastase dried in a similar manner, 30 parts bran flour, 5 parts wheat germs, and 25 to 20 parts of dried milk. The schizo-saccharomyces bomb may be cultivated in known manner in a dilute beer wort free from hops, 5 per cent raw sugar with additional nutrients, or grape must culture medium.  The cultivation may be at 30 DEG  C. with aeration, and the organism may be separated from the medium, washed and dried as such or after dilution.  For the drying the pulp is supplied in a thin layer to a heated rotary drum with a stripper blade at one side, the vapour being withdrawn by a vacuum pump.  Instead of drying the pure yeast, the culture medium containing it may be dried.  The drying kills the cells, and inactivates the zymase and enzymes which break down low-molecular albuminous